The invention relates to nucleic acids and polypeptides.
The Wnt gene family includes at least ten genes that encode structurally related secreted glycoproteins. Members of the Wnt family are reported to be regulators of mammary cell growth and differentiation. For example, members have been shown to control a variety of developmental processes, e.g., cell fate specification, cell proliferation, cell polarity and cell migration. In addition, dysregulation of Wnt signaling has been reported to cause developmental defects, and to be implicated in the genesis of several human cancers.
Wnt family gene members Wnt-4, -7b, and -11 are expressed in metanephric kidneys. The expression of members of the Wnt gene family has been examined during kidney formation and in processes associated with complete urethral obstruction. Urethral obstruction induced during metanephrogenesis disrupts the normal pattern of Wnt-7b expression and interferes with the normal transformation process in developing kidneys, by maintaining the mesenchymal component and inducing the transformation of epithelium to mesenchyme. Accordingly, Wnt proteins may serve as mediators of the transformation of renal mesenchyme to epithelium.
Overexpression of Wnt-7B proteins can result in cellular transformation of C57MG cells. Higher levels expression of Wnt7b has also been reported in superficial bladder cancer cells as compared to invasive bladder cancer. These results suggest the Wnt-7B protein is involved in the early events of bladder tumorigenesis.
The present invention is based, in part, upon the discovery of a novel human nucleic acid sequence encoding a polypeptide having sequence similarity to Wnt-7B. The Wnt-7B like nucleic acids, polynucleotides, proteins and polypeptides or fragments thereof described herein include those found in clone 29518614.0.61 (SEQ ID NO:1), and the polypeptide encoded by clone 29518614.0.61 (SEQ ID NO:2).
In one aspect, the invention includes an isolated Wnt-7B-like nucleic acid molecule which includes a nucleotide sequence encoding a polypeptide that includes the amino acid sequence of SEQ ID NO:2. For example, in various embodiments, the nucleic acid can include a nucleotide sequence that includes SEQ ID NO:1. Alternatively, the encoded Wnt-7B-like polypeptide may have a variant amino acid sequence, e.g., have an identity or similarity less than 100% to the disclosed amino acid sequences, as described herein.
The invention also includes an isolated polypeptide that includes the amino acid sequence of SEQ ID NO:2 or a fragment having at least 6 amino acids of these amino acid sequences. Also included is a naturally occurring polypeptide variant of a Wnt-7B-like polypeptide, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes under stringent conditions to a nucleic acid molecule consisting of a Wnt-7B-like nucleic acid molecule.
Also included in the invention is an antibody that selectively binds to a Wnt-7B-like polypeptide. The antibody is preferably a monoclonal antibody, and most preferably is a human antibody. Such antibodies are useful, for example, in the treatment of a pathological state in a subject wherein the treatment includes administering the antibody to the subject.
The invention further includes a method for producing a Wnt-7B-like polypeptide by culturing a host cell expressing one of the herein described Wnt-7B-like nucleic acids under conditions in which the nucleic acid molecule is expressed.
The invention also includes methods for detecting the presence of a Wnt-7B-like polypeptide or nucleic acid in a sample from a mammal, e.g., a human, by contacting a sample from the mammal with an antibody which selectively binds to one of the herein described polypeptides, and detecting the formation of reaction complexes including the antibody and the polypeptide in the sample. Detecting the formation of complexes in the sample indicates the presence of the polypeptide in the sample.
The invention further includes a method for detecting or diagnosing the presence of a disease, e.g., a pathological condition, associated with altered levels of a polypeptide having an amino acid sequence at least 80% identical to a Wnt-7B-like polypeptide in a sample. The method includes measuring the level of the polypeptide in a biological sample from the mammalian subject, e.g., a human, and comparing the level detected to a level of the polypeptide present in normal subjects, or in the same subject at a different time, e.g., prior to onset of a condition. An increase or decrease in the level of the polypeptide as compared to normal levels indicates a disease condition.
Also included in the invention is a method of detecting the presence of a Wnt-7B-like nucleic acid molecule in a sample from a mammal, e.g., a human. The method includes contacting the sample with a nucleic acid probe or primer which selectively hybridizes to the nucleic acid molecule and determining whether the nucleic acid probe or primer binds to a nucleic acid molecule in the sample. Binding of the nucleic acid probe or primer indicates the nucleic acid molecule is present in the sample.
The invention further includes a method for detecting or diagnosing the presence of a disease associated with altered levels of a Wnt-7B-like nucleic acid in a sample from a mammal, e.g., a human. The method includes measuring the level of the nucleic acid in a biological sample from the mammalian subject and comparing the level detected to a level of the nucleic acid present in normal subjects, or in the same subject at a different time. An increase or decrease in the level of the nucleic acid as compared to normal levels indicates a disease condition.
The invention also includes a method of treating a pathological state in a mammal, e.g., a human, by administering to the subject a Wnt-7B-like polypeptide to the subject in an amount sufficient to alleviate the pathological condition. The polypeptide has an amino acid sequence at least 80% identical to a Wnt-7B-like polypeptide.
Alternatively, the mammal may be treated by administering an antibody as herein described in an amount sufficient to alleviate the pathological condition.
Pathological states for which the methods of treatment of the invention are envisioned include a cancer, e.g., colorectal carcinoma, a prostate cancer benign tumor, an immune disorder, an immune deficiency, an autoimmune disease, acquired immune deficiency syndrome, transplant rejection, allergy, an infection by a pathological organism or agent, an inflammatory disorder, arthritis, a hematopoietic disorder, a skin disorder, atherosclerosis, restenosis, a neurological disease, Alzheimer""s disease, trauma, a surgical or traumatic wound, a spinal cord injury, and a skeletal disorder.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.
Other features and advantages of the invention will be apparent from the following detailed description and claims.